W 2800.7 E56s No.E-5 1989/90 c.3 SECTION 6 ENDANGERED SPECIES ACT OKLAHOMA o FEDERAL AID PROJECT E-5 SURVEY AND SPECIES DETERMINATION OF CAVE CRAYFISH IN OKLAHOMA JUNE 1, 1989 - MAY 31, 1990 This study was undertaken to determine the identification, distribution and abundance of troglobitic crayfish in the Ozark Plateau region of Oklahoma. Specific objectives were: 1. To survey sites with historical records of cave crayfish in the Ozarks of Oklahoma, and determine the current status of cave crayfish at those sites. 2. To survey other potential sites for cave crayfish for which no records of occurrence exist. 3. To determine which species are in which caves using tissue electrophoresis to clarify genetic similarity of populations. Information from this study is expected to be useful in making decisions on listing and protection needs for cave crayfish in the Ozark Plateau of Oklahoma, Missouri and Arkansas. Cave crayfish are troglobitic, entirely dependent upon a subterranean existence. While a divergent group taxonomically, they possess convergent characteristics of deceased pigmentation and claw musculature. They are represented in the Americas by species in eight families, which also have epigean species (Hobbs et. al. 1977). Four species have been named from the western part of the Ozark Plateau, which includes Oklahoma, Arkansas and Missouri. All are members of the subgenus Jugicambarus in the genus Cambarus (Hobbs and Brown 1987). These are considered to have derived from an epigean parental stock in the southern Appalachian Mountains (Hobbs et. al. 1977, Hobbs and Barr 1960). The only troglobitic species from the subgenus Jugicambarus that is known to occur outside of the Ozark Plateau is Cambarus cryptodytes from the panhandle of northern Florida (Hobbs and Barr 1960). The species Cambarus hubricti is a troglobitic species of a different subgenus that occurs in the eastern part of the Ozark Plateau (Gardner 1986, Hobbs et. al. 1977, Hobbs and Barr 1960).None of the troglobitic species from the Ozarks are known to co-occur. The main morphological feature used to distinguish the troglobitic species of Jugicambarus is the gonopod (first pleopod) of males in reproductive condition, called first form males (in this report abbreviated Male-I). While non-sexual characters can be used for tentative identifications, confirmed identifications must be based on the gonopods of Male-Is (Hobbs et. al. 1977, Hobbs 1976 and personal communication). The status of cave crayfish in Oklahoma is particularly problematic, principaUy because adequate data have not been coUected. Of the four species of the subgenus Jugicambarus described from the Ozark Plateau, two are thought to occur in Oklahoma. Cambarus setosus Faxon is known from eight counties in southwestern Missouri (Figg and Lister 1990). Prior to the mid 1970s, it was thought that C. setosus probably occurred in Oklahoma (Black 1971, Creaser and Ortenburger 1933, Williams 1954). On the basis of specimens that were not Male-I from Twin Cave and "Spring Creek, 5 mi. S. of Locust Grove" in Oklahoma, Hobbs and Barr (1960) tentatively caUed populations from those caves C. setosus. Cambarus tartarus Hobbs and Cooper was described from type specimens from a single cave in Oklahoma (Hobbs and Cooper 1972). Without having specimens to examine from three other Oklahoma caves (Jail, Rodman, and Star), the cave crayfish from those caves were tentatively assigned to C. tartarus (Hobbs et. al. 1977). Cambarus aculabrum Hobbs and Brown has been described from two caves in Arkansas, both of which are close to the Oklahoma state line (Hobbs and Brown 1987). Cambarus zophonastes Hobbs and Bedinger is known from a single cave in Stone County, Arkansas (Hobbs and Bedinger 1964), considerably to the east of Oklahoma and aU"other known sites for species of Jugicambarus (Figg and Lister 1990, Hobbs and Brown 1987, Hobbs et. al 1977). Prior to this study, cave surveys in the Ozark Plateau of Oklahoma had not focused on status assessment of cave crayfish, so records have been scattered and incomplete. Cambarus setosus: none Cambarus tartarus: Candidate 2, petitioned in 1986 for listing as endangered (Brown 1986). Candidate 2, petitioned for listing as endangered. Endangered, Federal Register for April 7, 1987. As troglobites, cave crayfish are obligate inhabitants of subterranean aquatic systems. In the Ozark Plateau of Oklahoma, some caves have sizeable perennial streams that follow the main course of the cave for its length; some caves have perennial streams and/or pools in only a part of the cave; and some caves have ephemeral streams and/or pools that have water only when water levels are raised by rains. Based on a few records on microhabitat use in Oklahoma caves, it appears that the cave crayfish are found more often in pools and lower gradient parts of streams. In all aquatic caves, water levels and gradient fluctuate with flood conditions. In most caves and subterranean aquatic systems, the lack of primary producers results in dependency of primary consumers and detritivores on the import of organic material from surface areas. Thus, though caves in Oklahoma may not have cave crayfish at all times, they may be important habitat with respect to sources for nutrient inputs. The study area was in that portion of the Ozark Plateau of Oklahoma in which aquatic caves are known to occur. This includes parts of Delaware, Mayes and Ottawa counties, principally where the surface geology is limestone and chert (Figures 1 and 2). Personal communication with Dr. Arthur Brown, Dr. Jeffery Black and Mr. William Puckette, who have seen cave crayfish in Oklahoma caves, indicated that all known specimens deposited in a museum are in the collection of the U. S. National Museum. I visited that collection in March of 1990 and examined all specimens of cave crayfish from Oklahoma. In addition, published and unpublished records were obtained and reviewed. Historical records also were obtained for the Ozark cavefish, Amblyopsis rosae, on the assumption that habitat suitable for a troglobitic fish might be suitable for a troglobitic crayfish. Caves selected for field survey work included those with reported records of cave crayfish and cavefish and those aquatic caves in close proximity to caves with historical records. The rationale was that caves in proximity were most likely to be in the same hydrological system as caves with historical records. Aquatic caves with no records of cave crayfish or cavefish were not visited if they had had frequent checks by W. L. Puckette during winter bat surveys. Figure 2. Detail of region in Oklahoma with aquatic solution caves limestone (shaded area). CHEROKEE surveys were conducted with W. L. Puckette who is most familiar with caves in the Ozarks of Oklahoma. Accessible ephemeral and perennial aquatic areas of each cave were visited. Two caves were visited two times because water levels were extremely low during the first visit and did not afford adequate assessment of crayfish use of those caves. A third cave was revisited to collect a male for morphometric examination. Electrophoretic Analysis of Tissue Proteins: When cave crayfish were found in a cave, a claw (usually the left claw) was removed from individuals that possessed two claws if the individuals appeared healthy. Samples were sent on dry ice to the Missouri Department of Conservation for electrophoretic analysis of the claw muscle tissue. Dr. Jeffery Koppelman performed the analyses along with samples of cave crayfish from Ozark Plateau caves in Missouri. We were unsuccessful in obtaining samples from Arkansas caves, principally because the people in that state who are familiar with Ozark caves were not available to do the work. Because of the sensitivity of caves of the Ozark Plateau and extreme pressures for human visitation use, the precise location of caves we visited is not presented in this report. Instead, by agreement with the U. S. Fish & Wildlife Service (Steven Hensley, Ecological Services Office, Tulsa, Oklahoma, personal communication), the location of known caves is given to the nearest four sections in Appendix 1. Caves are identified by the same code system that has been used in studies of endangered bats in caves in the study area. A total of 16 caves were located and surveyed, one spring was visited, and we were unable to locate one cave for which an historical record exists. A summary of the results of both the historical record and field surveys is given in Table 1. The results are detailed in Appendix 1. Cave crayfish were found in four caves, coded as DL-38, DL-39, DL-74 and DL-91 (Table 2 and Appendix I). Morphological measurements taken on live individuals were not an adequate means of distinguishing the species (Table 2 and H. H. Hobbs, Jr., personal communication). Males in reproductive condition (Male-I) were distinguished from non-reproductive males (Male-II) and examined for species determination. One Male-I was captured, examined and released in each of caves DL- 38, DL-74, and DL-91. Although the gonopods were viewed under a lOX hand lens, a positive determination of species could not be made. On the basis of published descriptions, Cambarus setosus can be distinguished from C. tartarus by a notch on the distal end of the central projection of the gonopod; the notch is lacking in C. setosus. No notch was seen on the gonopod of the Male-Is from DL-38, DL-74 or DL-91. Because of the apparent low density of these cave populations, no voucher specimens Summary of historical records and results